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      Characterization and Pathogenicity of Lasiodiplodia theobromae Causing Black Root Rot and Identification of Novel Sources of Resistance in Mulberry Collections

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          Abstract

          Black root rot (BRR) caused by Lasiodiplodia theobromae is an alarming disease of mulberry that causes tremendous economic losses to sericulture farmers in India and China. Successful control of this disease can be attained by screening germplasm and identifying resistant sources. Seventy four diseased root samples were collected from farmer’s fields belonging to four major mulberry growing states of South India. Based on morpho-cultural and scanning electron microscopy studies, 57 fungal isolates were characterized and identified as L. theobromae. Phylogenetic analysis of concatenated internal transcribed spacer and β-tubulin sequences revealed variation of the representative 20 isolates of L. theobromae. Following the root dip method of inoculation, pathogenicity studies on susceptible mulberry genotypes (Victory-1 and Thailand male) recognized the virulent isolate MRR-142. Accordingly, MRR-142 isolate was used to evaluate resistance on a set of 45 diverse mulberry accessions. In the repeated experiments, the mulberry accession ME-0168 which is an Indonesian origin belonging to Morus latifolia was found to be highly resistant consistently against BRR. Eight accessions (G2, ME-0006, ME-0011, ME-0093, MI-0006, MI-0291, MI-0489, and MI-0501) were found to be resistant. These promising resistant resources may be exploited in mulberry breeding for developing BRR resistant varieties and to develop mapping populations which successively helps in the identification of molecular markers associated with BRR.

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          MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms.

          The Molecular Evolutionary Genetics Analysis (Mega) software implements many analytical methods and tools for phylogenomics and phylomedicine. Here, we report a transformation of Mega to enable cross-platform use on Microsoft Windows and Linux operating systems. Mega X does not require virtualization or emulation software and provides a uniform user experience across platforms. Mega X has additionally been upgraded to use multiple computing cores for many molecular evolutionary analyses. Mega X is available in two interfaces (graphical and command line) and can be downloaded from www.megasoftware.net free of charge.
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            AMPLIFICATION AND DIRECT SEQUENCING OF FUNGAL RIBOSOMAL RNA GENES FOR PHYLOGENETICS

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              Development of primer sets designed for use with the PCR to amplify conserved genes from filamentous ascomycetes.

              We constructed nine sets of oligonucleotide primers on the basis of the results of DNA hybridization of cloned genes from Neurospora crassa and Aspergillus nidulans to the genomes of select filamentous ascomycetes and deuteromycetes (with filamentous ascomycete affiliations). Nine sets of primers were designed to amplify segments of DNA that span one or more introns in conserved genes. PCR DNA amplification with the nine primer sets with genomic DNA from ascomycetes, deuteromycetes, basidiomycetes, and plants revealed that five of the primer sets amplified a product only from DNA of the filamentous ascomycetes and deuteromycetes. The five primer sets were constructed from the N. crassa genes for histone 3, histone 4, beta-tubulin, and the plasma membrane ATPase. With these five primer sets, polymorphisms were observed in both the size of and restriction enzyme sites in the amplified products from the filamentous ascomycetes. The primer sets described here may provide useful tools for phylogenetic studies and genome analyses in filamentous ascomycetes and deuteromycetes (with ascomycete affiliations), as well as for the rapid differentiation of fungal species by PCR.
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                Author and article information

                Journal
                Plant Pathol J
                Plant Pathol J
                The Plant Pathology Journal
                Korean Society of Plant Pathology
                1598-2254
                2093-9280
                August 2022
                1 August 2022
                : 38
                : 4
                : 272-286
                Affiliations
                [1 ]Molecular Biology Laboratory-1, Central Sericultural Research and Training Institute, Mysuru 570 008, Karnataka, India
                [2 ]Department of Genetics and Plant Breeding, Sampurna International Institute of Agri Science & Horticultural Technology, Mandya 571 433, Karnataka, India
                [3 ]Department of Microbiology, Bhavan’s Vivekananda College of Science, Humanities and Commerce, Secunderabad 500 094, Telangana, India
                Author notes
                [* ]Corresponding author: Phone) +91-7893900859, FAX) +91-821-2362845, E-mail) gnaneshbn@ 123456gmail.com

                Handling Editor: Junhyun Jeon

                Author information
                https://orcid.org/0000-0003-0253-5394
                Article
                ppj-oa-01-2022-0005
                10.5423/PPJ.OA.01.2022.0005
                9372095
                35953047
                990fe05a-4049-4b21-b1b4-4d6e96796868
                © The Korean Society of Plant Pathology

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 20 January 2022
                : 6 April 2022
                : 2 May 2022
                Categories
                Research Article

                black root rot,lasiodiplodia theobromae,mulberry,pathogenicity,resistance

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